Cardiovascular disease is the leading cause of death in the United States. Systemic arterial hypertension (or simply "hypertension") is a major risk factor for the development of cardiovascular disease. Aldosterone and desoxycorticosterone (DOC) are mineralocorticoids that cause hypertension through increased urinary sodium (Na) retention and hypokalemic metabolic alkalosis through greater urinary acidification. Mineralocorticoids affect distal nephron and collecting duct (CD) ion transport to achieve these effects. The CD expresses at least two isoforms of H,K-ATPases, HK?1 and HK?2, that participate in acid secretion by the CD. Our recent studies also show that chronic DOC pivalate (DOCP) administration stimulates renal Na retention, weight gain, and metabolic alkalosis that are dependent on the presence of H,K-ATPases. Specifically, mice that lack both H,KATPases (HK?1,2 -/-) failed to develop significant Na retention or metabolic alkalosis in response to DOCP stimulation. Comparison of the response of the single HK?1 -/- to the double HK?1,2 -/- strongly suggests an important physiological role for the H,K-ATPase HK?2 isoform in the response to DOCP. Therefore, our central hypotheses are that: A) the H,K-ATPase HK?2 isoform is critical for the chronic action of mineralocorticoids to produce renal Na retention, cause hypokalemic metabolic alkalosis, and increase blood pressure (BP);and that B) dietary K loading abolishes or attenuates mineralocorticoid action on Na retention and electrolyte abnormalities, which are dependent on the induction of the H,K-ATPase HK?2 isoform in the kidney. We propose the following specific aims: 1) to determine the role of the H,K-ATPase HK?2 isoform in the ion transport response to chronic mineralocorticoid treatment;2) to determine the role of H,K-ATPase HK?2 isoform in the regulation of blood pressure and the contribution of K intake and hypokalemia to these changes;3) to determine if the differences in WT and HK?2 KO animals in response to DOCP administration reflect a renal phenotype. We will test the novel physiological hypothesis that the H,K-ATPase HK?2 isoform has an important role in Na homeostasis, volume regulation and BP control. As such, we have developed an integrated approach that combines whole animal metabolic balance studies, in vitro microperfusion, state-ofthe- art radiotelemetry measurement of BP, and renal transplantation to examine the physiological significance of HK?2 stimulation by mineralocorticoids.
These studies have direct relevance to our understanding of electrolyte disorders such as hypokalemia and metabolic alkalosis, which are potentially lethal disorders that are frequently seen in the veteran population. In addition, this research will provide a better understanding of the fundamental mechanisms of sodium regulation by the kidney. Excessive sodium absorption mediated by aldosterone and mineralocorticoids predictably leads to hypertension that is prevalent in the veteran population. Hypertension is a major cause of death among veterans due to the development of cardiovascular disease and the high incidence of stroke in hypertensive patients. These diseases are the leading cause of death among veterans as well as the general population of the United States. These studies examine a novel mechanism for renal sodium retention by mineralocorticoids and aldosterone. This proposal should identify new targets for drug therapy and intervention to more effectively control renal sodium regulation and hypertension that is so prevalent in the veteran population.